Evidence for DNA Polymerase‐α and ‐β Activity in Sugar Beet

Tymonko, John M.; Dunham, Valgene L.

doi:10.1111/j.1399-3054.1977.tb01487.x

Cited by 23 publications

(11 citation statements)

References 19 publications

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“…DNA polymerases which share some, but not all, of the properties of the animal and yeast a-type DNA polymerases have been reported from cauli¯ower (Fukasawa et al 1980), maize (Coello et al 1992;Coello-CoutinÄ o et al 1994;Coello and VaÂ zquez-Ramos 1995), pea (Chivers and Bryant 1983;Bryant et al 1992), periwinkle (Gardner and Kado 1976), rice (Amileni et al 1979), spinach (Misumi and Weissbach 1982), sugar beet (Tymonko and Dunham 1977), turnip (Dunham and Bryant 1986) and wheat (Castroviejo et al 1979;Graveline et al 1984;Laquel et al 1990aLaquel et al ,b, 1994. Only a few of these studies have reported on associated primase activities.…”

Section: Introductionmentioning

confidence: 94%

Purification and properties of a DNA primase from Nicotianatabacum

García-Maya

Buck

1997

Planta

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A DNA primase was isolated from a nuclear fraction from leaves of tobacco (Nicotiana tabacum L. cv. Samsun) and from purified nuclei prepared from tobacco suspension culture cells. The DNA primase was purified to homogeneity (i) for preparations from leaves, by ammonium sulphate fractionation, followed by chromatography on columns of phosphocellulose, Q-Sepharose, heparin-Sepharose and single-stranded DNA cellulose, and sedimentation in a glycerol gradient, or (ii) for preparations from cells, by chromatography on single-stranded DNA cellulose, followed by ammonium sulphate precipitation and chromatography on columns of High Q, heparin-Sepharose and Mono Q. In glycerol gradients, the DNA primase sedimented at a rate corresponding to a molecular mass of about 120 kDa. In SDS-polyacrylamide gel electrophoresis, the primase was resolved into two polypeptide subunits of 63 kDa and 53 kDa, which are similar in size to the primase subunits of animal and yeast DNA polymerase alpha-primase complexes. On poly(dT) or phage M13 single-stranded DNA templates, the DNA primase catalysed the synthesis of oligoribonucleotides up to 20 nucleotides in length, which could serve as primers for DNA synthesis catalysed by Escherichia coli DNA polymerase. Primase activity was dependent on a template, magnesium ions and ATP; it was resistant to aphidicolin and rifampicin, but was strongly inhibited by N-ethylmaleimide. This is the first report of the purification to homogeneity of a plant DNA primase.

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Section: Introductionmentioning

confidence: 94%

Purification and properties of a DNA primase from Nicotianatabacum

García-Maya

Buck

1997

Planta

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“…Crude enzyme preparations or partially purified enzymes have been used. Examples of e-type (replicative) are: sugar beet [32], pea [9,29], wheat [71, spinach [24], cauliflower [14] and rice [1 ]. fl-type enzymes (repair) have been described for wheat [8], cauliflower [ 14] and soybean [ 10].…”

Section: Introductionmentioning

confidence: 99%

A DNA polymerase from maize axes: its purification and possible role

Coello¹,

Rodrı́guez²,

García³

et al. 1992

Plant Mol Biol

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Three different DNA polymerase activities can be resolved by passing a protein extract from 24 h imbibed maize axes through DEAE-cellulose. These activities have been numbered 1, 2 and 3, according to their elution order. One of them, DNA polymerase 2, elutes at 100-120 mM phosphates. This enzyme was further purified by passing it through Heparin-Sepharose, Sephacryl S-300 and DNA cellulose. Purification was nearly 5000-fold. The enzyme needs Mg2+, is stimulated by K+, has an optimum pH of 7.0 and its optimum temperature is 30-37 degrees C. Specific inhibitors for different types of polymerases, such as aphidicolin, dideoxythymidine triphosphate and N-ethyl maleimide, gave intermediate values of inhibition, making impossible the definition of the type of enzyme purified by its inhibitory pattern. SDS-PAGE indicated the presence of several bands of molecular masses of 28-40, 56 and 15 kDa. Most of these bands could be visualized when proteins from crude extracts were analyzed by western blot, using an antibody against calf thymus DNA polymerase alpha. A high molecular mass (around 500 kDa) was calculated by western blot of native gels using the same antibody. Finally, specific activity of this enzyme increased 100-fold during maize germination whereas polymerase 3 virtually did not increase. Furthermore, immunoprecipitation experiments with the antipolymerase alpha-antibody showed a decrease in DNA polymerase activity by 70%. The possibility that polymerase 2 is a replicative enzyme is discussed.

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“…that they do not possess a DNA polymerase-/?-like enzyme, but that they do possess one or more enzymes which have some similarities to polymerase-a and polymerase-y. Other investigators (Srivastava, 1974; Tymonko & Dunham, 1977; have suggested that higher plants contain DNA polymerases of direct equivalence to the DNA polymerases-a, -/?, -y of vertebrates. Table 3 shows that many higher plants, covering a relatively wide taxonomic range, have been shown to possess a DNA polymerase which has some properties in common with the DNA polymerase-a.…”

Section: ( D ) Dna Polymerases Of Higher Plantsmentioning

confidence: 99%

Biochemical Aspects of Dna Replication With Particular Reference to Plants

Bryant¹

1980

Biological Reviews

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Evidence for DNA Polymerase‐α and ‐β Activity in Sugar Beet

Cited by 23 publications

References 19 publications

Purification and properties of a DNA primase from Nicotianatabacum

Purification and properties of a DNA primase from Nicotianatabacum

A DNA polymerase from maize axes: its purification and possible role

Biochemical Aspects of Dna Replication With Particular Reference to Plants

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